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The role of crustins in Litopenaeus vannamei in response to infection with shrimp pathogens: An in vivo approach
Authors:Jessica E Shockey  Nuala A O’Leary  Enrique de la Vega  Craig L Browdy  John E Baatz  Paul S Gross
Institution:1. Marine Biomedicine and Environmental Sciences Center, Medical University of South Carolina, 221 Fort Johnson Road, Charleston, SC 29412, USA;2. Marine Resources Institute, South Carolina Department of Natural Resources, 217 Fort Johnson Road, Charleston, SC 29412, USA;3. Department of Pediatrics, Medical University of South Carolina, DCRI, MSC 513, 173 Ashley Ave., Charleston, SC 29425, USA;4. Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC 29425, USA
Abstract:Crustin antimicrobial peptides, identified in crustaceans, are hypothesized to have both antimicrobial and protease inhibitor activity based on their primary structure and in vitro assays. In this study, a reverse genetic approach was utilized to test the hypothesis that crustins are antimicrobial in vivo in response to bacterial and fungal challenge. Injection of double-stranded RNA specific to a 120-bp region of LvABP1, one of the most prominent crustin isoforms, yielded a significant reduction in the expression of both crustin mRNA and protein within the hemocytes. To test the role of crustins in the shrimp immune response, RNAi was first used to suppress crustin expression and animals were subsequently injected with low pathogenic doses of either Vibrio penaeicida or Fusarium oxysporum. A significant increase in mortality in crustin-depleted animals was observed in animals infected with V. penaeicida as compared to controls, whereas no significant change in shrimp mortality was observed following infection with F. oxysporum.
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